title: Generation of an ICF syndrome model by efficient genome editing of human induced pluripotent stem cells using the CRISPR system.
abstract: Genome manipulation of human induced pluripotent stem (iPS) cells is essential to achieve their full potential as tools for regenerative medicine. To date, however, gene targeting in human pluripotent stem cells (hPSCs) has proven to be extremely difficult. Recently, an efficient genome manipulation technology using the RNA-guided DNase Cas9, the clustered regularly interspaced short palindromic repeats (CRISPR) system, has been developed. Here we report the efficient generation of an iPS cell model for immunodeficiency, centromeric region instability, facial anomalies syndrome (ICF) syndrome using the CRISPR system. We obtained iPS cells with mutations in both alleles of DNA methyltransferase 3B (DNMT3B) in 63% of transfected clones. Our data suggest that the CRISPR system is highly efficient and useful for genome engineering of human iPS cells.
authors.affiliation: Laboratory of Genome Science, Biosignal Genome Resource Center, Institute for Molecular and Cellular Regulation, Gunma University, Gunma 371-8512, Japan. hatada@gunma-u.ac.jp.
authors.forename: Takuro
authors.initials: T
authors.lastname: Horii
authors.forename: Daiki
authors.initials: D
authors.lastname: Tamura
authors.forename: Sumiyo
authors.initials: S
authors.lastname: Morita
authors.forename: Mika
authors.initials: M
authors.lastname: Kimura
authors.forename: Izuho
authors.initials: I
authors.lastname: Hatada
mesh_terms: D000483:Alleles; D001483:Base Sequence; D002460:Cell Line; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics; D018390:Gene Targeting / Q000379:methods; D005818:Genetic Engineering / Q000379:methods; D016678:Genome / Q000235:genetics*; D006801:Humans; D006972:Hypertelorism / Q000235:genetics*; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism*; D008607:Intellectual Disability / Q000235:genetics*; D007738:Kyphosis / Q000235:genetics*; D058627:Megalencephaly / Q000235:genetics*; D008969:Molecular Sequence Data; D017393:RNA Editing / Q000235:genetics*; D014059:Tongue / Q000002:abnormalities*; D000097584:DNA Methyltransferase 3B
chemical_list: D004248:DNA (Cytosine-5-)-Methyltransferases; D000097584:DNA Methyltransferase 3B